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1、棉花学报CottonScience2016,28(3):283―290响应面法对解淀粉芽孢杆菌()12-7产抗菌蛋白条件的优化1211*王全袁王占利袁高同国袁李术娜渊1.河北农业大学生命科学学院袁河北保定071001曰2.保定学院资源与环境系袁河北保定071051冤摘要院解淀粉芽孢杆菌()12-7是从发病棉田土壤中分离得到的1株生防细菌袁其对棉花黄萎病致病菌大丽轮枝菌Kleb.V190具有明显的拮抗作用袁已证明其抗菌物质为蛋白质遥采用响应面法对该生防菌株的产抗菌蛋白条件进行优化袁以期为棉花黄萎病生物农药的研制开发以及抗菌蛋白的后续研究奠定基础遥以发酵液
2、上清液对大丽轮枝菌的抑菌圈面积为考察指标袁首先采用单因子试验考察培养时间尧温度尧初始pH尧碳源尧氮源尧无机盐等因素对菌株12-7发酵产抗菌蛋白的影响曰再根据响应面分析法的基本原理和方法袁对发酵条件进行优化遥得出该菌株产抗菌蛋白的最优发酵培养基和培养条件为院1.5%渊质量分数冤蔗糖尧1.5%渊质量分数冤大豆蛋白胨尧0.3%渊质量分数冤NaCl尧0.1%渊质量分数冤K2HPO4尧初始pH7.3的培养基袁31℃发酵培养60h遥优化后抑菌圈面积从试验设计初期的171.95mm2提高至320.3mm2袁且经过验证试验证实预测值与实际值之间具有较好的拟合度遥关键词
3、院响应面法曰大丽轮枝菌曰拮抗细菌曰抗菌蛋白曰发酵条件曰优化中图分类号院S482.7文献标志码院A文章编号院1002-7807渊2016冤03-0283-0810.11963/issn.1002-7807.2016030121211*WangQuan,WangZhanli,GaoTongguo,LiShuna(1.071001,;2.071000,)Inapreviousstudy,thenovelstrain12-7,whichhasahighantifungalactivityagainstKleb.,wasisolatedandscreenedfr
4、omthesoilofadiseasedcottonfield.Itwasdemonstratedthatantifungalmaterialsproducedby12-7wereproteins.Inthisstudy,responsesurfacemethodology(RSM)wasusedtooptimizeantagonisticproteinfermentationparameters.Thisworklaysthefoundationforthedevelopmentofantifungalproteinbiologicalsaspest
5、i-cides,andforfollow-upstudies.Takingtheinhibitionzoneareainducedbythefermentationsupernatantasstandard,singlefac-torialexperimentsweredesignedtodetermineeffectsofincubationtime,temperature,incipientpHvalue,differentcarbonandnitrogensources,andinorganicsaltontheantifungalactivit
6、yofthe12-7strain.Then,RSMwasusedtooptimizethefermenta-tionparameters.Thisrevealedtheoptimumfermentationofstrain12-7toproduceantagonisticproteinwasasfollows:mediumwith1.5%(massfraction)sucrose,1.5%soypeptone,0.3%NaCl,0.1%K2HPO4,andincipientpHvalue7.3;withcultivationat31℃for60h.An
7、tifungalactivityvalueincreasedfrom171.95mm2intheinitialexperimentaldesignto320.3mm2undertheoptimumcultivationcondition.Theexperimentaldataundervariousconditionsthusvalidatedthetheoreticalvalues.responsesurfacemethodology;Kleb.;antagonisticbacterium;antagonisticprotein;fermentati
8、oncondition;optimization收稿日期:2015-01-22作者简介院王全渊
